The present study was undertaken to evaluate and compare the in vitro leucoagglutination titers and in vivo lymphopenic eftect of three different antilymphocytic sera. Rabbits were immunized against adult dog and pup lymphoid tissue with multiple intramuscular injections of pooled spleen, mesenteric lymphnode and thymus cells. The number of lymphoid cells per each immunization was 1.8 * 107 of spleen cells, 1.68*108 of mesenteric lymphnode cells and 4.4*107 of thymus cells and immunization was done once per 10 days for 2 months. After completion of immunization and one booster injection, the rabbits were bled and total of 11 antidog spleen sera(RADSS), 4 antidog lymphnode sera(RADLS)and 3 antidog thymus sera (RADTS) were prepared and stored at-20C until used. With in vitro agglutination technique, each serum was serially diluted with buffered saline and tested with freshly prepared dog spleen, lymphnode,thymus cells and peripheral blood buffy coat leucocytes. With in vivo test, the antilymphocytic sera were intramuscularly injected in a dose of 1 ml/kg into adult dogs and after 12 hours peripheral blood pictures were evaluated and compared with those before antisera administration. The results were as follows. 1. The range of in vitro leucoagglutination titers of RADSS for spleen cells, tymphnode cells, thymus cells and peripheral leucocytes was 1: 4 to 1: 128, 1: 4 to 1: 128, 1: 4 to 1: 64 and 1: 4 to 1: 128, respectively. 2. The range of in vitro leucoagglutination titers of RADLS for sp]een cells, lymphnode cells, thymus cells and peripheral leucocytes was 1: 32 to 1: 64. 1: 32 to 1: 256, 1: 32 to 1: 128 and 1: 16 to 1: 256, respectively. 3. The range of in vitro leucoagglutination titers of RADTS for spleen cells, lymphnode cells, thymus cells and peripheral leucocytes was 1: 16 to l: 128, 1: 16 to 1: 64, 1: 8 to 1: 64 and 1: 8 to 1: 64, respectively. 4. After a singIe intramuscular injection of the antisera, the total R.B,C. counts were not significantly changed in RADSS, RADLS and RADTS. However, the percentage of neutrophils increased bv a mean value of 35.3% 34.8% and 21.7%p in RADSS,RADI.S and RADTS, respectively and the percentage of lymphocytes decreased by a mean value of 47.8%, 51.8% and 30.6% in RADSS, RADLS and RADTS, respectiveIy. The eosinophils showed a tendency of decrease in RADSS, RADLS and RADTS, respectively. The monocyes and basophis were not significantly changed after injection of the antisera. The author found that there was no significant difference between the in vitro leucoagglutination titers of three kind of antisera and that with in vitro leucoagglutination test, the peripheral blood leucocytes may serve as an useful antigen because of simp]icity of preparation. Also, the author found that there was no direct correlation between the in vitro leucoagglutination titers and in vivo Iymphopenic effect, but generally the antisera with strong lymphopenic effect had higher in vitro titers than those with weak lymphopenic effcct, In this experiment, the lymphopenic effect of RADTS was slightly lawer than those ot RADSS and RADLS and this difference may be due to smaller number of the cells used in immunization for the preparation of the antisera. The author concluded that spleen was as useful as lymphnode or thymus in its antigenicity and in producing effective antilymphocytic serum.
Adult ; Agglutination ; Animals ; Blood Buffy Coat ; Dogs ; Eosinophils ; Humans ; Immune Sera ; Immunization ; Injections, Intramuscular ; Lymphocytes ; Lymphoid Tissue ; Neutrophils ; Rabbits ; Spleen* ; Thymus Gland*

buffy coat, fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) from 130 samples.RESULTS: The rate of repeated tests in a total of buffy coat, FF and FFPE was 0%, 8%, and 34%, respectively. The accuracy of BRCA1/2 NGS testing was 100.0%, 99.9% and 99.9% in buffy coat, FFPE and FF, respectively. However, due to the presence of variant allele frequency (VAF) shifted heterozygous variants, tumor materials (FFPE and FF) showed lower sensitivity (95.5%–99.0%) than buffy coat (100%). Furthermore, FFPE showed 51.4% of the positive predictive value (PPV) on account of sequence artifacts. When performed in the post-filtration process, PPV was increased by approximately 20% in FFPE. Buffy coat showed 100% of sensitivity, specificity and accuracy in BRCA1/2 NGS test.CONCLUSIONS: On the comparison of the analytical performance according to different sample types, the buffy coat was not affected by sequencing artifacts and VAF shifted variants. Therefore, the blood test should be given priority in detecting germline BRCA1/2 mutation, and tumor materials could be suitable to detect somatic mutations in OC patients without identifying germline BRCA1/2 mutation.]]>
Artifacts ; Blood Buffy Coat ; Gene Frequency ; Hematologic Tests ; High-Throughput Nucleotide Sequencing ; Humans ; Mass Screening ; Ovarian Neoplasms ; Sensitivity and Specificity ; Tissue Preservation

BACKGROUND: The concept of "neovascularization", which was first applied to describe the pathogenesis of diseases such as diabetic retinopathy or rheumatoid arthritis, has been extended to other fields of study such as myocardial ischemia and tumorigenesis. Endothelial progenitor cells (EPCs), play a critical role in neovascularization and have been reported to also be capable of colonizing vascular grafts. In this study, EPCs were isolated from cord blood, peripheral blood and bone marrow, and then cultured. Various cytokines, such as vascular endothelial growth factor(VEGF), Insulin growth factor(IGF), endothelial growth factor(EGF) fibroblast growth factor-basic(FGF-b), stem cell factor(SCF), flt3-ligand(FL), and thrombopoietin(TPO) were added to the cultures and observed for their effects on endothelial cells for their potential use in antineoplastic therapy or treatment of regional ischemia. METHODS: The mononuclear cells (MNCs) were isolated from cord blood, peripheral blood buffy coat, and bone marrow. They were collected from healthy donors using Ficoll-Hypaque. CD34+ cells were isolated by MACS system. To evaluate the effect of various cytokines, purified CD34+ cells were cultured under conditions of various cytokine combinations including SCF, Fl, TPO, VEGF, EGF, IGF, and FGF-b. After four weeks of culture, umbilical cord blood and bone-marrow derived adherent cells were analyzed for endothelial markers by immunohistochemical stain. RESULTS: Cultured adherent cells expressed the endothelial specific markers, such as KDR, CD34, CD31, CD62E, and CD cadherin but did not express vWF antigen. Typical morphology of endothelial cells was observed, such as the cord-like structure and cobblestone appearance during the culture period, which suggested that the adherent cells were consistent with endothelial cells. CONCLUSION: We described the experimental conditions in which endothelial progenitors were differentiated from CD34+ cells isolated from three hematopoietic stem cell sources: bone marrow, peripheral blood and cord blood.
Arthritis, Rheumatoid ; Blood Buffy Coat ; Bone Marrow ; Bone Marrow Cells ; Carcinogenesis ; Colon ; Cytokines ; Diabetic Retinopathy ; Endothelial Cells ; Epidermal Growth Factor ; Fetal Blood ; Fibroblasts ; Hematopoietic Stem Cells* ; Humans ; Insulin ; Ischemia ; Myocardial Ischemia ; Stem Cells* ; Tissue Donors ; Transplants ; Vascular Endothelial Growth Factor A